February 17, 2014 FOR SUSPECTED SALMONELLA AND SHIGELLA COLONIES (gram-negative rods) Subcultivate in Himedia SS (Salmonella-Shigella) Agar

1. Suspend __ grams in __ ml distilled water. 2. Boil with frequent agitation to dissolve the medium completely. 3. DO NOT AUTOCLAVE OR OVERHEAT. Overheating may destroy selectivity of the medium. 4. Cool to about 50C. 5. Mix and pour into sterile Petri plates. *Quality Control Appearance: Light yellow to pink homogeneous free flowing powder Colour and Clarity of prepared medium: Reddish orange coloured clear to slightly opalescent gel forms in Petri plates FOR SUSPECTED STAPHYLOCOCCUS COLONIES (gram-positive cocci) Subcultivate in Himedia Mannitol Salt Agar Base 1. Suspend __ grams in __ ml distilled water. 2. Heat to boiling to dissolve the medium completely. 3. Sterilize by autoclaving at 15 lbs pressure (121C) for 15 minutes. 4. If desired, add 5% v/v Egg Yolk Emulsion (FD045). 5. Mix well and pour into sterile Petri plates. *Quality Control Appearance: Light yellow to pink homogeneous free flowing powder Colour and Clarity of prepared medium: Red coloured clear to slightly opalescent gel forms in Petri plates FOR SUSPECTED E. COLI COLONIES (gram-negative rods) Subcultivate in BBL Eosin Methylene Blue Agar 1. Suspend __ g of the powder in __ mL of purified water. Mix thoroughly. 2. Heat with frequent agitation and boil for 1 minute to completely dissolve the powder. 3. Autoclave at 121C for 15 minutes. 4. Cool to approximately 45C. Agitate gently and pour into plates. *Quality Control Dehydrated Appearance: Fine, homogeneous, may contain up to a large amount of minute to small dark red purple particles Prepared Appearance: Medium to dark, green orange brown, hazy OR Subcultivate in MacConkey Agar with 0.15% Bile salts, CV and NaCl 1. Suspend __ grams in __ ml distilled water. 2. Heat to boiling with gentle swirling to dissolve the agar completely. 3. Sterilize by autoclaving at 15 lbs pressure (121C) for 15 minutes. Avoid overheating. 4. Cool to 45 - 50C and pour into sterile Petri plates. The surface of the medium should be dry when inoculated. *Quality Control Appearance: Light yellow to pink homogeneous free flowing powder Colour and Clarity of prepared medium: Red with purplish tinge coloured clear to slightly opalescent gel forms in Petri plates.